Anti-KV1.5 (KCNA5)-ATTO-550 Antibody

Potassium voltage-gated channel subfamily A member 5
    Cat #: APC-004-AO
  • Lyophilized Powder
  • Antigen Incl.
  • Shipped at Room Temp.
  • Type: Polyclonal
    Source: Rabbit
    Reactivity: h, m, r
    Immunogen
    GST fusion protein with sequence HRETDHEEQAALKEEQGIQRRESGLDTGGQRKVSCSKASFHKTGGPLEST DSIRRGSCPLEKCHLKAKSNVDLRRSLYALCLDTS RETDL, corresponding to amino acid residues 513-602 of mouse KV1.5 (Accession Q61762), (MW: 37 kDa.). Intracellular, C-terminus.
    Accession (Uniprot) Number Q61762
    Gene ID 16493
    Peptide confirmation Confirmed by DNA sequence and SDS-PAGE.
    Homology Rat - 86/90 amino acid residues identical; rabbit - 71/90 amino acid residues identical; human - 70/90 amino acid residues identical; bovine, dog - 66/90 amino acid residues identical.
    Purity The serum was depleted of anti-GST antibodies by affinity chromatography on immobilized GST and from antibodies cross-reactive to other KV1 by affinity chromatography on immobilized KV1.3-GST fusion proteins, and then the antibody was affinity purified on immobilized KV1.5-GST.
    Formulation Lyophilized powder. Reconstituted antibody contains phosphate buffered saline (PBS), pH 7.4, 1% BSA, 5% sucrose, 0.05% NaN3.
    Label ATTO-550. Maximum absorption 554 nm; maximum fluorescence 576 nm. The fluorescence is excited most efficiently in the 540 - 565 nm range. This label is related to the dye Rhodamine 6G and can be used with filters used to detect Rhodamine.
    Storage before reconstitution The antibody ships as a lyophilized powder at room temperature. Upon arrival, it should be stored at -20°C.
    Reconstitution 50 µl double distilled water (DDW).
    Antibody concentration after reconstitution 1 mg/ml.
    Storage after reconstitution The reconstituted solution can be stored at 4°C, protected from the light, for up to 1 week. For longer periods, small aliquots should be stored at -20°C. Avoid multiple freezing and thawing. Centrifuge all antibody preparations before use (10000 × g 5 min).
    Control antigen storage before reconstitution Lyophilized powder can be stored intact at room temperature for 2 weeks. For longer periods, it should be stored at -20°C.
    Control antigen reconstitution 100 μl PBS.
    Control antigen storage after reconstitution -20°C.
    Preadsorption Control 3 μg fusion protein per 1 μg antibody.
    Standard quality control of each lot Western blot analysis (unlabeled antibody, #APC-004), and immunohistochemistry (labeled antibody).
    Applications: ih
    May also work in: ic
    Immunohistochemistry
    Expression of KV1.5 in mouse cerebellum
    Immunohistochemical staining of perfusion fixed, free-floating frozen mouse brain sections using Anti-KV1.5 (KCNA5)-ATTO-550 Antibody (#APC-004-AO), (1:50), (red). Staining was detected in cerebellar Bergmann glial cells (arrows). DAPI is used as the counterstain (blue). G = granule layer, P = Purkinje layer, M = molecular layer.
    Expression of KV1.5 in rat cerebellum
    Immunohistochemical staining of perfusion fixed, free-floating frozen rat brain sections using Anti-KV1.5 (KCNA5)-ATTO-550 Antibody (#APC-004-AO), (1:50), (red). Staining was detected in cerebellar Bergmann glial cells (arrows). DAPI is used as the counterstain (blue). G = granule layer, P = Purkinje layer, M = molecular layer.
    References
    1. Swanson, R. et al. (1990) Neuron 4, 929.
    2. Gutman, G.A. et al. (2005) Pharmacol. Rev57, 473.
    3. McGahon, M.K. et al. (2007) Am. J. Physiol. 292, H1001.
    Scientific background

    KV1.5 is a mammalian voltage dependent K+ channel, homologous to the Drosophila Shaker K+ channel. KV1.5 was first cloned from the rat brain.1 Eight Shaker related genes exist in mammals constituting the KV1, subfamily of the large KV channel family of genes.2

    A functional KV1 channel is either a membrane spanning homotetramer or heterotetramer, which is composed of members of the same subfamily. In addition several auxiliary subunits and intracellular proteins might interact with the channel and affect its function.

    The structure of KV1.5 channel is similar to all KV channels and includes six membrane spanning helices creating a voltage sensor domain and a pore domain.2

    The channel is expressed in cardiac and smooth muscle tissue (colon, aorta, stomach and pulmonary artery) as well as in neurons and kidney.2 A loss-of-function mutation in the gene encoding the channel was found in atrial fibrillation patients, stressing its role as a cardiac action potential regulator.3

    The functional channel is considered transient (A-type) channel and shows prominent inactivation. Therefore, this channel activity influences the membrane potential and excitability of neurons and muscle.

    KV1.5 channels are sensitive to high doses of TEA (330 mM) and low doses of 4-AP (0.27 mM), the “classical” non-selective potassium channel blockers.2

    Application key:

    CBE- Cell-based ELISA, FC- Flow cytometry, ICC- Immunocytochemistry, IE- Indirect ELISA, IFC- Indirect flow cytometry, IHC- Immunohistochemistry, IP- Immunoprecipitation, LCI- Live cell imaging, N- Neutralization, WB- Western blot

    Species reactivity key:

    H- Human, M- Mouse, R- Rat
    Image & Title:

    Anti-Kv1.5 (KCNA5)-ATTO-550 Antibody
    Immuno-colocalization of KV1.3 and KV1.5 in mouse cerebellum
    Immunohistochemical staining of mouse perfusion-fixed frozen brain sections using Anti-KV1.3 (KCNA3) (extracellular)-Biotin Antibody (#APC-101-B) (1:400), and Anti-KV1.5 (KCNA5)-ATTO-550 Antibody (#APC-004-AO), (1:60). A. KV1.3 staining (green) is detected in Bergmann glia soma and processes in the molecular layer (Mol) (arrow). B. Same section shows staining for KV1.5 (red). C. Merge of the two images suggests considerable co-localization in the soma of Bergmann glia. In the molecular layer, the distribution of KV1.5 is diffuse, unlike the discrete staining for KV1.3 in glial processes. Cell nuclei were visualized with DAPI (blue).

    Last update: 27/11/2018

    Anti-KV1.5 (KCNA5) Antibody (#APC-004) is a highly specific antibody directed against an epitope of the mouse protein. The antibody can be used in western blot, immunohistochemistry, immunocytochemistry, and immunoprecipitation applications. It has been designed to recognize KV1.5 from human, rat, and mouse samples.

    Anti-KV1.5 (KCNA5)-ATTO-550 Antibody (#APC-004-AO) is directly labeled with an ATTO-550 fluorescent dye. ATTO dyes are characterized by strong absorption (high extinction coefficient), high fluorescence quantum yield, and high photo-stability. The ATTO-550 fluorescent label is related to the well known dye Rhodamine 6G and can be used with filters typically used to detect Rhodamine. Anti-KV1.5 (KCNA5)-ATTO-550 Antibody is specially suited to experiments requiring simultaneous labeling of different markers.

    For research purposes only, not for human use
    Citations
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